Container with film sparger

ABSTRACT

A container assembly includes a flexible bag having an interior surface bounding a chamber and an opposing exterior surface, the bag having a top end wall, a bottom end wall, and an encircling sidewall extending therebetween, the bottom end wall including a first polymeric sheet overlying a second polymeric sheet. A first weld line welds the first polymeric sheet to the second polymeric sheet, the first weld line bounding a first sparging area formed between the first polymeric sheet and the second polymeric sheet. A plurality of first perforations are formed through a portion of the first polymeric sheet that overlies the first sparging area so that gas can pass from the first sparging area, through the first perforations and into the chamber of the bag.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit to Provisional Application No.61/541,913, filed Sep. 30, 2011, which is incorporated herein byspecific reference.

BACKGROUND OF THE INVENTION

1. The Field of the Invention

The present invention relates to spargers incorporated into a flexiblebag.

2. The Relevant Technology

Spargers are commonly used in bioreactors for delivering controlledvolumes of gas to a growth media containing cells. In part, the gas isused to control the partial pressure of oxygen within the growth mediaand to control the pH and other perimeters of the growth media so thatthe conditions are optimal for cell growth. Spargers typically comprisea hollow metal ring having a hose coupled thereto. The ring is formedfrom a sintered metal so that the ring is porous. The ring is manuallypositioned at the bottom of a container with the hose extending upthrough a port at the top of the container. During operation,pressurized gas is delivered to the ring through the hose. The gas thenpermeates out through the metal ring so as to enter the media in theform of small bubbles. As the bubbles travel up through the media, atleast a portion of the gas becomes entrained within the media. Otherconventional spargers comprise a section of stainless steel tubing thatis bent into a ring with small diameter holes positioned along thecurved length thereof

Although conventional spargers are useful in delivering gas to themedia, they have a number of shortcomings. For example, conventionalspargers are relatively expensive to make and are thus designed to bereused. Reuse of a conventional sparger, however, requires that it beremoved from the container and then cleaned and sterilized. In somesituations, cleaning of the sparger can be difficult in that cellby-product, dead cells, and other particulate within the growth mediacan be lodged on or trapped within the sparger. Thus cleaning andsterilizing of the sparger can be both time consuming and expensive.Time and care must also be taken to properly position and seal thesparger within the container without contaminating the sparger or thecontainer.

Furthermore, in conventional bioreactors it is necessary that the growthmedia containing the cells be continually mixed or suspended so that theproperties of the growth media remain homogeneous. Conventional spargerscan obstruct the flow of the fluid which can produce dead spots wherethe cells die. Furthermore, the cells can be caught on or by the spargerwhich can damage or kill the cells. In addition, the spargers must becarefully designed and positioned so that they do not obstruct themixing system.

Some current bioreactors comprise a flexible bag that is disposed withina rigid support housing. The cell culture is grown within the sterilecompartment of the flexible bag. In an attempt to eliminate some of theabove sparger problems, disposable spargers have been incorporated intothe flexible bags. Such disposable spargers comprise a port having anenlarged annular flange welded to the inside of the bag and a tubularstem that projects from the flange to outside the bag. The stem bounds apassage the extends through the flange porous film overlays the flangeinside of the bag so as to cover the passage and is welded around theperimeter edge of the flange. As a result, gas can be passed through thestem from outside the bag. The gas passes through the flange and thenpasses through the porous film where it enters the cell culture withinthe bag in the form of small bubbles. When the cell production iscompleted, the bag and associated sparger are simply disposed of.

Although the above flexible sparger eliminates some of the problems ofconventional spargers, the new bag spargers also have theirshortcomings. Most notably, the bag spargers only sparge at a relativelysmall, fixed location on the bag and are limited to only one size of gasbubbles. As such, bag spargers have limited or no adjustability withregard to sparging at different locations, flow rates, bubbles sizes, orcombinations of the forgoing.

Accordingly, what is needed are spargers and container systems that cansolve one or more of the above shortcomings.

BRIEF DESCRIPTION OF THE DRAWINGS

Various embodiments of the present invention will now be discussed withreference to the appended drawings. It is appreciated that thesedrawings depict only typical embodiments of the invention and aretherefore not to be considered limiting of its scope.

FIG. 1 is a cross section view of one embodiment of a container assemblywithin a support housing, a container assembly incorporating a sparger;

FIG. 2 is an exploded perspective view of the bottom end wall of thecontainer assembly shown in FIG. 1;

FIG. 3 is a top plan view of the bottom end wall of the containerassembly shown in FIG. 1 showing the spargers formed thereon and amanifold coupled thereto;

FIG. 4 is a top plan view of the container assembly shown in FIG. 3having an alternative manifold coupled thereto;

FIG. 5 is a top plan view of the bottom end wall shown in FIG. 3 havinga plurality of discrete gas lines coupled thereto;

FIG. 6 is a top plan view of a bottom end wall of a container assemblyhaving an alternative configuration of spargers formed thereon;

FIG. 7 is a top plan view of a bottom end wall of a container having analternative embodiment of a sparger mounted thereon;

FIG. 8 is an exploded perspective view of a bottom end wall of acontainer containing three sheets;

FIG. 9 is a cross sectional side view of an alternative embodiment of acontainer assembly having spargers extending down through the bottom endwall;

FIG. 10 is a top plan view of the bottom end wall of the container shownin FIG. 9;

FIG. 11 is a cross sectional side view of one of the spargers shown inFIG. 9;

FIG. 12 is a perspective view of the floor of the support housing shownin FIG. 9; and

FIG. 13 is a cross sectional side view of an alternative embodiment of asparger where a gas line projects from the sparger up into thecontainer.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention relates film spargers as well as container systemsthat incorporate such spargers. In general, one embodiment of a filmsparger comprises overlying sheets of flexible material wherein one ormore weld lines weld the sheets together so that a sparging area isbounded between the overlying sheets. A gas line is in communicationwith the sparging area for delivering a gas thereto while perforationsare formed through one of the sheets so that gas passing into thesparging area can pass out through the perforations for sparging afluid. Film spargers are typically incorporated into a flexible bag orother type of container for sparging a fluid within the container or forotherwise delivering gas bubbles to the fluid within the container.

Depicted in FIG. 1 is one embodiment of a containment system 10incorporating features of the present invention. Containment system 10comprises a substantially rigid support housing 12 in which a containerassembly 13 is disposed. Support housing 12 has an upper end 14, a lowerend 16, and an interior surface 18 that bounds a compartment 20. Formedat lower end 16 is a floor 22. An encircling sidewall 23 extends up fromfloor 22 toward upper end 14. As will be discussed below in greaterdetail, one or more openings 24 can extend through floor 22 or sidewall23 of support housing 12 so as to communicate with compartment 20. Upperend 14 terminates at a lip 26 that bounds an inlet opening 28 tocompartment 20. If desired, a cover, not shown, can be mounted on upperend 14 so as to cover inlet opening 28. Likewise, an access opening canbe formed at another location on support housing 12 such as throughsidewall 12 at second end 16 or through floor 22. The access opening islarge enough so that an operator can reach through the access opening tohelp manipulate and position container assembly 13. The access openingcan be selectively closed by a door or cover plate.

It is appreciated that support housing 12 can come in a variety ofdifferent sizes, shapes, and configurations. For example, floor 22 canbe flat, frustoconical, or have other slopes. Sidewall 23 can have atransverse cross section that is circular, polygonal or have otherconfigurations. Support housing 24 can be insulated and/or jacketed sothat a heated or cooled fluid can flow through the jacket for heating orcooling the fluid contained within container assembly 13. Compartment 20can be any desired volume such as those discussed below with regard tocontainer 32.

As also depicted in FIG. 1, container assembly 13 is at least partiallydisposed within compartment 20 of support housing 12. Container assembly13 comprises a container 32 having one or more ports 52 mounted thereon.In the embodiment depicted, container 32 comprises a flexible bag havingan interior surface 38 that bounds a chamber 40 suitable for holding afluid 41 or other type of material. More specifically, container 32comprises a side wall 42 that, when container 32 is inflated, can have asubstantially circular or polygonal transverse cross section thatextends between a first end 44 and an opposing second end 46. First end44 terminates at a top end wall 48 while second end 46 terminates at abottom end wall 50.

Container 32 can be comprised of one or more sheets of a flexible, waterimpermeable material such as a low-density polyethylene or otherpolymeric sheets having a thickness typically in a range between about0.1 mm to about 5 mm with about 0.2 mm to about 2 mm being more common.Other thicknesses can also be used. The material can be comprised of asingle ply material or can comprise two or more layers that are eithersealed together or separated to form a double wall container. Where thelayers are sealed together, the material can comprise a laminated orextruded material. The laminated material can comprise two or moreseparately formed layers that are subsequently secured together by anadhesive.

The extruded material can comprise a single integral sheet thatcomprises two or more layers of different material that are eachseparated by a contact layer. All of the layers are simultaneouslyco-extruded. One example of an extruded material that can be used in thepresent invention is the HyQ CX3-9 film available from HyCloneLaboratories, Inc. out of Logan, Utah. The HyQ CX3-9 film is athree-layer, 9 mil cast film produced in a cGMP facility. The outerlayer is a polyester elastomer coextruded with an ultra-low densitypolyethylene product contact layer. Another example of an extrudedmaterial that can be used in the present invention is the HyQ CX5-14cast film also available from HyClone Laboratories, Inc. The HyQ CX5-14cast film comprises a polyester elastomer outer layer, an ultra-lowdensity polyethylene contact layer, and an EVOH barrier layer disposedtherebetween. In still another example, a multi-web film produced fromthree independent webs of blown film can be used. The two inner webs areeach a 4 mil monolayer polyethylene film (which is referred to byHyClone as the HyQ BM1 film) while the outer barrier web is a 5.5 milthick 6-layer coextrusion film (which is referred to by HyClone as theHyQ BX6 film).

The material can be approved for direct contact with living cells and becapable of maintaining a solution sterile. In such an embodiment, thematerial can also be sterilizable such as by ionizing radiation.Examples of materials that can be used in different situations aredisclosed in U.S. Pat. No. 6,083,587 that issued on Jul. 4, 2000 andU.S. Patent Publication No. US 2003/0077466 A1, published Apr. 24, 2003that are each hereby incorporated by specific reference.

In one embodiment, container 32 comprises a two-dimensional pillow stylebag wherein two sheets of material are placed in overlapping relationand the two sheets are bounded together at their peripheries to forminternal chamber 40. Alternatively, a single sheet of material can befolded over and seamed around the periphery to form internal chamber 40.In another embodiment, container 32 can be formed from a continuoustubular extrusion of polymeric material that is cut to length and theends seamed closed.

In still other embodiments, container 32 can comprise athree-dimensional bag that not only has an annular side wall but also atwo-dimensional top end wall 48 and a two-dimensional bottom end wall50. For example, three-dimensional container 32 can comprise sidewall 42formed from a continuous tubular extrusion of polymeric material that iscut to length, such as shown in FIG. 2. A circular top end wall 48(FIG. 1) and bottom end wall 50 can then be welded to opposing ends ofsidewall 42. In yet another embodiment, three-dimensional container 32can be comprised of a plurality of discrete panels, typically three ormore, and more commonly between four to six. Each panel can besubstantially identical and comprises a portion of side wall 42, top endwall 48, and bottom end wall 50 of container 32. The perimeter edges ofadjacent panels are seamed together to form container 32. The seams aretypically formed using methods known in the art such as heat energies,RF energies, sonics, or other sealing energies. In alternativeembodiments, the panels can be formed in a variety of differentpatterns.

It is appreciated that container 32 can be manufactured to havevirtually any desired size, shape, and configuration. For example,container 32 can be formed having chamber 40 sized to 10 liters, 30liters, 100 liters, 250 liters, 500 liters, 750 liters, 1,000 liters,1,500 liters, 3,000 liters, 5,000 liters, 10,000 liters or other desiredvolumes. Chamber 40 can also have a volume in a range between about 10liters to about 5,000 liters or about 30 liters to about 1,000 liters.Any other ranges selected from the above volumes can also be used.Although container 32 can be any shape, in one embodiment container 32is specifically configured to be complementary to or substantiallycomplementary to compartment 20 of support housing 12.

In any embodiment, however, it is typically desirable that whencontainer 32 is received within compartment 20, container 32 isgenerally uniformly supported by support housing 12. Having at leastgenerally uniform support of container 32 by support housing 12 helps topreclude failure of container 32 by hydraulic forces applied tocontainer 32 when filled with fluid.

Although in the above discussed embodiment container 32 is in the formof a flexible bag, in alternative embodiments it is appreciated thatcontainer 32 can comprise any form of collapsible container, flexiblecontainer, or semi-rigid container. Furthermore, in contrast to having aclosed top end wall 48, container 32 can comprise an open top liner.Container 32 can also be transparent or opaque and can have ultravioletlight inhibitors incorporated therein.

Mounted on top end wall 48 are a plurality of ports 52 that are in fluidcommunication with chamber 40. Although two ports 52 are shown, it isappreciated that one or three or more ports 52 can be present dependingon the intended use of container 32. As such, each port 52 can serve adifferent purpose depending on the type processing to be undertaken. Forexample, ports 52 can be coupled with a tube 54 for dispensing fluid orundertaken. ; other components into chamber 40 or withdrawing fluid fromchamber 40. In addition, such as when container 32 is used as abioreactor for growing cells or microorganisms, ports 52 can be used toprovide various probes, such as temperature probes, pH probes, dissolvedoxygen probes, and the like, access to chamber 40. It is appreciatedthat ports 52 can come in a variety of different configurations and canbe placed at any number of different locations on container 32,including sidewall 42 and bottom end wall 50.

Although not required, in one embodiment means are provided for mixingfluid 41 within chamber 40. The means for mixing can be in the form of amixing assembly. By way of example and not by limitation, in oneembodiment as shown in FIG. 1 a drive shaft 56 projects into chamber 40and has an impeller 58 mounted on the end thereof. A dynamic seal 59forms a seal between shaft 56 and container 32. External rotation ofdrive shaft 56 facilitates rotation of impeller 58 that mixes and/orsuspends fluid 41 within chamber 40. Specific examples of how toincorporate a rotational mixing assembly into a flexible container aredisclosed in U.S. Pat. No. 7,384,783 that issued Jun. 10, 2008 and U.S.Pat. No. 7,682,067 that issued on Mar. 23, 2010, which are incorporatedherein by specific reference.

In yet another alternative embodiment of the means for mixing or themixing assembly, mixing can be accomplished by vertically reciprocallymoving a vertical mixer within chamber 40. Further disclosure withregard to the assembly and operation of vertical mixer is disclosed inU.S. Patent Publication No. 2006/0196501, published Sep. 7, 2006, whichis incorporated herein by specific reference. In yet other embodiments,it is appreciated that the mixing can be accomplished by simplycirculating fluid through chamber 40 such as by using a peristaltic pumpto move fluid in and out of chamber 40; by rotating a magnetic impelleror stir bar within container 32 and/or by injecting sufficient gasbubbles within the fluid to mix the fluid. Other conventional mixingtechniques can also be used.

Continuing with FIG. 1, bottom end wall 50 has a plurality of spargersincorporated therein. Specifically, bottom end wall 50 comprises a firstsheet 60 having a first side face 62 and an opposing second side face64. First sheet 60 overlays a second sheet 66 that likewise has a firstside face 68 and an opposing second side face 70. First sheet 60 andsecond sheet 66 typically comprise flexible polymeric sheets such asthose discussed above with regard to container 32. As discussed abovewith regard to bottom end wall 50, first sheet 60 can comprise acontinuous sheet that is welded to side wall 42 around a perimeter edge69 as depicted in FIG. 2. Alternatively, first sheet 60 can comprise anintegral portion of sidewall 42 or can comprise a plurality of separatesheets secured together that are either attached to or are an integralportion of sidewall 42. Second sheet 66 can be welded to second sideface 64 of first sheet 60 and/or welded to sidewall 42, such as along aperimeter edge 71 of second sheet 66. In other embodiments, second sheet66 can be welded to or comprise an integral portion of sidewall 42, asdiscussed above with regard to first sheet 60, while first sheet 60 iswelded or otherwise secured to first side face 68 of second sheet 66and/or sidewall 42.

Depicted in FIG. 3 is a top plan view of first sheet 60 overlayingsecond sheet 66. In this embodiment, sheets 60 and 66 are weldedtogether by a weld line 72. Weld line 72, as with other weld linesdiscussed herein, can be formed using any conventional technique such aslaser welding, sonic welding, heat welding, or the like. Weld line 72 isshown as welding together the perimeter or outside edges of sheets 60and 66 but can be formed radially inward from one or both of theperimeter edges or at other locations. As also shown in FIG. 3, fourseparate spargers 74A-D are formed by producing other weld lines betweensheets 60 and 66.

For example, sparger 74A is formed by forming a weld line 76A starts ata first location 78A located at or adjacent to the perimeter edge ofsheet 60 and/or sheet 66 and extends into the interior of sheets 60 and66 along a predetermined path for the sparger 74A and then circles backto a second location 80A at or adjacent to the perimeter edge of sheet60 and/or sheet 66 adjacent to first location 78A. Weld line 76A boundsa perimeter of a sparger pathway 82 a which is the area bounded betweensheets 60 and 66 and partially encircled by weld line 76A. In theembodiment depicted, sparger pathway 82A comprises a gas transfer path84A that extends from a first end 86 to an opposing second end 88. Anopening 87A is formed at first end 86 between locations 78A and 80A andbetween sheets 60 and 66 through which a gas can be fed into gastransfer path 84A. Sparger pathway 82A also comprises a sparging area90A formed at second end 88 that is in fluid communication with gastransfer path 84A. In the embodiment depicted, gas transfer path 84 is anarrow elongated path while sparging area 90 forms an enlarged circulararea. Other configurations can also be used.

A plurality of perforations 92 extend through first sheet 60 of spargingarea 90A so that gas can pass along gas transfer path 84A, into spargingarea 90A and then out through perforation 92 to form gas bubbles withinfluid 41 disposed within chamber 40. Spargers 74B-D are similarly formedwith like reference characters being used to identify like elements. Byusing this technique, a plurality of discrete spargers can be easilyformed on container 32. Each sparger can be disposed at any desiredlocation and be any desired size, shape or configuration. Likewise,although four spargers 82 are shown, it is appreciated that any numberof spargers such as 1, 2, 3, 5, or more can be formed with sheets 60 and66. The sparging areas can be uniformly distributed over sheets 60 and66 or can be located at defined locations for optimal sparging. Forexample, a sparger can be disposed directly below the means for mixingsuch that the mixing or movement of fluid 41 produced by the mixer helpsto entrain the gas bubbles within fluid 41.

In some embodiments, each sparger can have the same number ofperforations 92 and all perforations 92 can be the same size and shape.In alternative embodiments, perforations 92 can be different between twoor more different spargers. For example, different spargers can havedifferent numbers, sizes, and/or shapes of perforations 92 to optimizeperformance in different situations. Larger perforations 92 producelarger gas bubbles that may be optimal for striping CO₂ from fluid 41whereas smaller perforations produce smaller bubbles that may bepreferred for oxygenating fluid 41. Likewise, increasing the number ofperforations 92 may be helpful in causing the bubbles to mix the fluidand/or increase stripping or oxygenation. In other embodiments, it isappreciated that one or more of spargers 74A-D can have combinations ofdifferent perforations 92. For example, a single sparger can have bothsmall and large perforations 92. In one embodiment, the smaller bubblesare formed from perforations 92 typically having a diameter of less than0.8 mm, 0.4 mm or 0.2 mm, 0.1 mm while the large bubbles are formed fromperforation typically having a diameter greater than 1.5 mm, 0.8 mm, 0.4mm or 0.15 mm. Perforations of other diameters can also be used. Thesize of the perforation and resulting bubbles depends on the intendeduse and the size of container 32. For example, the large bubbles aretypically larger when processing a large volume of fluid in a largecontainer than when processing a relatively small volume of fluid in asmall container. The variance or delta between the diameter of theperforations for the small bubbles and the perforations for the largebubbles is typically at least 0.15 mm, 0.3 mm, 0.5 mm or 1 mm and isoften within ±0.1 mm or ±0.5 of these values. Other variances can alsobe used.

As discussed below in greater detail, spargers 74A-D can simultaneouslyoperate or, alternatively, a manifold or other regulator can be used sothat one or more of the spargers can be operated while the otherspargers are not operated. Accordingly, by having different spargerswith different perforations 92, select spargers can be used in differentsituations or times to optimize performance.

In some embodiments, it is appreciated that gas transfer path 84A ofsparger 74A is not required. For example, perforations 92 can be formedthrough first sheet 60 overlying gas transfer path 84A so as to convertgas transfer path 84A in a portion of sparging area 90A. It isappreciated that perforations 92 can be formed using any conventionaltechniques. For example, perforations 92 can be formed as part of themanufacturing process for the sheet or can be subsequently produced bypunches or other techniques. In one embodiment, one or more lasers canbe used to form perforations 92. An advantage of using a laser is thatperforations 92 can be formed at precise locations and with a precisediameter so that bubbles can be formed having a precise, predefinedsize. Furthermore, when a laser is used to form a perforation, thematerial melted by the laser gathers around the perimeter edge of theperforation, thereby reinforcing the perforation and helping to preventrupture of the sheet.

In one embodiment of the present invention, a manifold can be used forcontrolling the gas flow to one or more of spargers 74A-D. For example,depicted in FIG. 3 is one embodiment of a manifold 100 incorporatingfeatures of the present invention. Manifold 100 comprises a body 102having a gas inlet port 104 and a plurality of gas outlet ports 106A-D.Gas outlet ports 106A-D are in parallel communication with gas inletport 104 by way of a forked flow path 108. A gas source, such as acompressor or a canister of compressed gas, is fluid coupled with gasinlet port 104. The gas can be air, oxygen, or any other gas orcombination of gases. Gas lines 110A-D extend from gas outlet ports106A-D, respectively, to a corresponding opening 87A-D at first end 86of each sparger 74A-D, respectively. Gas lines 110A-D can be weldedbetween sheets 60 and 66 at openings 87A-D so as to seal openings 87A-Dclosed. Gas lines 110A-D can comprise flexible or rigid tubes and can beintegrally formed with or separately attached to body 102.

Valves 112A-D are mounted on body 102 and control the flow of gas toeach gas line 110A-D, respectively. In one embodiment, valves 112A-D canbe electrical valves, such as solenoid valves, that can be used to open,close, or restrict the flow of gas to spargers 74A-D. In thisembodiment, electrical wiring 114 can couple to valves 112A-D forcontrolling their operation. In other embodiments, valves 112A-D cancomprise valves that are operated manually, hydraulically,pneumatically, or otherwise. By using manifold 100, different spargersor different combinations of spargers can be used at different times tooptimize performance as discussed above.

Depicted in FIG. 4 is an alternative embodiment of a manifold 100Awherein like elements between manifold 100 and 100A are identified bylike reference characters. Manifold 100A includes body 102 having gaslines 110A-D projecting therefrom and communicating with spargers 74A-D.In manifold 100A, gas lines 110A-D comprise flexible tubing. In turn,manifold 100A has valves 114A-D in the form of pinch clamps or hoseclamps that are mounted on gas lines 110A-D, respectively. It isappreciated that pinch clamps 114 can come in a variety of differentconfigurations and are used to manually pinch gas lines 110A-D so as tocontrol the flow of gas therethrough.

In other alternative embodiments, it is appreciated that a manifold isnot required. For example, as depicted in FIG. 5, gas lines 110A-D canextend from spargers 74A-D and have valves 114A-D coupled thereon,respectively. However, gas lines 110A-D need not be part of or coupledwith a manifold but rather can be separately coupled to discrete gassources if desired.

As previously discussed, any desired number, size, shape, and/orconfiguration of spargers can be formed. For example, depicted in FIG. 6is a top plan view of bottom end wall 50 having three spargers 118A-Cformed thereon. Again, the perimeter of each sparger 118A-C is formed byweld lines 120A-C, respectively, formed between sheets 60 and 66. Ifdesired, a single weld line can form a common boundary between adjacentspargers. For example, weld line 120B is shown forming a common boundarybetween spargers 118B and 118C. Sparger 120A is similar to sparger 74Aexpect that sparger 118A is more centrally located on sheets 60 and 66.Sparger 118B has a substantially C-shaped sparging area 122B that curvesaround sparging area 122A. Likewise, sparger 118C has a sparging area122C that substantially encircles around sparging area 122B.

In contrast to the prior spargers, sparger 118C has a first end 124 andan opposing second end 126 with gas lines 110A and 110D fluid coupledtherewith, respectively. In this configuration, a gas can be suppliedthrough both gas lines 110A and 110D at opposing ends of sparger 118C sothat the gas is more uniformly provided to sparging area 122C. As aresult, the gas exits out of all of perforations 92 at a more uniformpressure and flow rate. Again, the flow of gas into each of spargers118A-C can be controlled by the depicted manifold 100A or any other typeof manifold.

In the embodiment depicted in FIG. 6, impeller 58 (FIG. 1) can bevertically aligned with sparger 118A while sparger 118C is laterallyspaced apart from impeller 58. Sparger 118A can be designed to producesmall bubbles that interact with and are distributed by impeller 58throughout the fluid. By dispersing the small bubbles by using impeller58, the small bubbles have a longer dwell time within the fluid whichenhances mass transfer of the gas. For example, the bubbles can moreefficiently oxygenate the fluid. Sparger 118C produces larger bubblesthat do not directly interact with impeller 58. The larger bubbles arecommonly used for stripping CO₂ from the fluid. Because the largerbubbles have a higher buoyancy than the smaller bubbles, the impellerhas less of an influence on the larger bubbles and thus there may be noneed for aligning them with the impeller. Furthermore, the impeller maybreak up the larger bubbles making them less efficient for strippingCO₂. In addition, aligning the larger bubbles with the impeller cancause the impeller to cavitate which reduces mixing efficiency of thefluid. In other embodiments, however, sparger 118A can be designed toproduce large bubbles that are intentionally broken up and dispersed byimpeller 58 while sparger 118C produces small bubbles that do notdirectly interact with impeller 58. Other configuration can also beused.

In another embodiment, it may be desirable to have a single sparger thatcovers a large portion of bottom end wall 50 so that the fluid withinthe container can be more uniformally sparged. For example, depicted inFIG. 7 is a sparger 130 having a first end 134 and an opposing secondend 136 with a perimeter bounded by weld lines 132A and 132B extendingtherebetween. Sparger 130 is elongated and snakes along bottom end wall50 in a curving pattern. Sparger 130 has a sparging area 130, i.e., thearea between weld lines 132A and 132B, that covers at least 40% and morecommonly at least 50%, 60% or 80% of the surface area of one side ofbottom end wall 50. Other percentages can also be used. Gas lines 110Aand 110B are coupled with the opposing ends of sparger 130 so that a gascan be delivered to the opposing ends of sparger 130. As a result, thegas is more uniformly passed out through perforations 92 than if only asingle gas line was used.

Turning to FIG. 8, in one alternative embodiment the inventive spargerscan be formed by overlaying three or more sheets. For example, one ormore weld lines can weld sheet 60 to sheet 66 so as to form a sparger,represented by broken lines 141A, therebetween. Likewise, one or moreweld lines can weld sheets 66 and 140 together so as to form a sparger,represented by broken lines 141B, therebetween. An opening 142 can beformed through sheet 60 so as to expose the sparging area of sparger 141B formed between sheets 66 and 140. It is also appreciated that the weldlines can simultaneously weld all three sheets 16, 60, and 140 togetherto form sparger 141A and/ or 141B. Gas lines 110A and B couple to anddeliver gas to spargers 141A and 141B, respectively.

In the previous embodiments, the gas lines feeding gas to the spargersentered through an opening, such as openings 87A-D in FIG. 3, formedbetween sheets 60 and 66. This configuration enables the gas lines toproject radially out from the side of container 32 and thus to projectthrough an opening 24 (FIG. 1) in side of support housing 12. In analternative embodiment, however, the gas lines can fluid couple with thespargers so as to project down from the bottom of bottom end wall 50 andin turn project down through floor 22 of support housing 12. Forexample, as depicted in FIG. 9, bottom end wall 50 of container 32 isagain comprised of first sheet 60 overlapping second sheet 66 which arewelded together to form spargers 144A-C. Specifically, in the top planview as shown in FIG. 10, weld lines 146A-C weld sheets 60 and 66together and are each formed in a circular pattern so as to bound aperimeter of sparging areas 148A-C, respectively. In alternativeembodiment, it is appreciated that weld lines 146A-C can be formed inany encircling pattern. Spargers 144A-C also include perforations 92formed through first sheet 60 overlaying each sparging area 148.

As depicted in FIGS. 9 and 11, a port 152A has a flange 153 that ismounted on first side face 68 of second sheet 66 so that a stem 154extends down through an opening 156 in sheet 66. As a result, port 152Acommunicates with sparging area 148A of sparger 144A. Ports 152B and Care similarly coupled with spargers 144B and C. As shown in FIG. 9, afirst end of gas lines 110A-C is coupled with ports 152A-C,respectively, while an opposing second end of gas lines 110A-C iscoupled with manifold 100. As a result, manifold 100 can be used tocontrol selective operation of each of the spargers 144A-C. Again,spargers 144A-C can be of any desired size, shape or configuration.

Depending on the desired configuration for the containers and spargers,it is appreciated that the containers can be assembled using a varietyof different procedures. For example, either before or after cuttingsheets 60 and 66 to their desired size, perforations 92 can be formed onfirst sheet 60 having the desired number, size, shape and location.Likewise, where applicable, openings 156 can be formed on second sheet66 and ports 152 welded thereto. Next, sheets 60 and 66 can beoverlapped and the various weld lines formed so as to weld sheets 60 and66 together and produce the spargers. Where ports 156 are not used, gaslines can be welded within the opening formed between sheets 60 and 66so as to communicate with the spargers. Finally, sheets 60 and 66 can bewelded to side wall 42. Alternatively, sheet 60 can be welded to sidewall 42 and then second sheet 66 can be welded to first sheet 60 forwelding the sheets together and forming the spargers. In yet otherembodiments, first sheet 60 can be integrally formed with side wall 42or side wall 42 and first sheet 60 can comprise multiple sections thatare welded together. In these configurations, second sheet 66 wouldsubsequently be welded to the combination of first sheet 60 and sidewall 42. In other embodiments, second sheet 66 can be attached to or beintegrally formed with side wall 42 while first sheet 60 can comprise asmaller sheet or multiple smaller sheets that only cover a portionsecond sheet 66. Once the gas lines are coupled with different spargersand, where appropriate, a manifold coupled thereto, all gas lines andports coupled to the container are closed off and the full assembly issterilized by radiation or other traditional techniques.

To facilitate use, the container assembly 13 is lowered into compartment20 of support housing 12. The related manifold and/or gas lines are thenpassed out of compartment 20 through opening 24, when support housing 12shown in FIG. 1 is used. In the embodiment shown in FIG. 9, the manifoldand gas lines can pass down through an elongated opening 24A formed infloor 22 of support housing 12A. In both embodiments, a plate 158 can beused to help cover a portion of opening 24 or 24A after the manifold ispassed therethrough so as to minimize the size of the openings. Thiscovering of the openings reduced stress on the container caused by fluidtrying to push the container through the openings. Container assembly 13can be partially inflated with a gas after it is positioned withinsupport housing 12 to enable it to be manually adjusted and properlypositioned within support housing 12. Alternatively, container assembly13 can be filed with fluid while it is adjusted for proper positioning.Container assembly 13 can subsequently be used as a bioreactor,fermentor, or for simply processing fluids or chemicals.

In another alternative embodiment of the present invention, the gaslines can fluid couple with the spargers so as to project up from thetop of bottom end wall 50 and in turn couple with or extend out througha port 52 (FIG. 1) located at the upper end of container 32. Forexample, as depicted in FIG. 13, bottom end wall 50 of container 32 isagain comprised of first sheet 60 overlapping second sheet 66 which arewelded together to form a sparger 160. Specifically, in the same manneras previously discussed with regard to FIG. 10, a weld line 162 can weldsheets 60 and 66 together in a circular pattern so as to bound aperimeter of a sparging area 164. In alternative embodiments, it isappreciated that weld line 162 can be formed in any encircling patternand that any number of separate spargers 160 can be formed. It is againappreciated that first sheet 60 need only be large enough to formsparger 160 or multiple spargers 160 and need not be as large as secondsheet 66. Sparger 160 also includes perforations 92 formed through firstsheet 60 overlaying sparging area 164.

Flange 153 of port 152 is mounted on second side face 64 of first sheet60, such as by welding or adhesive, so that stem 154 extends out throughan opening in sheet 66. As a result, port 152 communicates with spargingarea 164. A first end 168 of a gas line 171 is coupled with a barbed end155 of port 152 while an opposing second end 172 of gas line 171 iseither coupled with or extends out through one of ports 52 (FIG. 1) ofcontainer 32. It is appreciated that port 52 to which gas line 171couples or extends out can be located at the upper end of container 32,as shown in FIG. 1, or can be located at any location along sidewall 42or on floor 50. One method for coupling second end 172 of gas line 171to port 52 is disclosed in U.S. Pat. No. 7,225,824, issued Jun. 5, 2007which is incorporated herein by specific reference. In turn, second end172 of gas line 171 can be placed in communication with a gas source fordelivering gas to sparger 160. Gas line 171 can also be coupled with amanifold which can control the flow of gas to multiple separate spargers160 formed on bottom end wall 50.

It is appreciated that the inventive spargers and related containershave a variety of unique advantages over conventional spargers. Forexample, the inventive spargers can be easily formed by simply weldingtwo sheets together. This welding can be achieved using the sameequipment and techniques used in forming the container. This ease inmanufacturing permits greater versatility in forming spargers of desiredsize, orientation, configuration, location, number and the like tooptimize desired processing parameters. Furthermore, the spargers areflexible and are part of the bag or container. This enables the combinedcontainer and spargers to be easily rolled up or folded withoutpotential risk of damage to the assembly. The rolled or folded assemblycan be easily sterilized, stored, shipped, and incorporated into a rigidsupport housing. The ability to produce multiple spargers at the bottomof the bag also enables spargers to be formed with differentperforations sizes so that different bubble sizes and numbers can beselectively produced to achieve different objectives. Furthermore, byusing the manifolds, operation of the different spargers or spargercombinations can be controlled to further optimize processingparameters. Because the combined container and spargers are relativelyinexpensive to make, the assembly can be designed as a disposable,single use item, thereby eliminating cleaning and sterilization betweenuses. The spargers are also flush with the floor so that they do notobstruct the flow of fluid or the flow of cells or microorganisms withinthe fluid.

The present invention may be embodied in other specific forms withoutdeparting from its spirit or essential characteristics. The describedembodiments are to be considered in all respects only as illustrativeand not restrictive. The scope of the invention is, therefore, indicatedby the appended claims rather than by the foregoing description. Allchanges which come within the meaning and range of equivalency of theclaims are to be embraced within their scope.

What is claimed is:
 1. A container assembly comprising: a flexible baghaving an interior surface bounding a chamber and an opposing exteriorsurface, the bag having a top end wall, a bottom end wall, and anencircling sidewall extending therebetween, the bottom end wall beingcomprised of a first polymeric sheet overlying a second polymeric sheet;an first weld line welding the first polymeric sheet to the secondpolymeric sheet, the first weld line bounding a first sparging areaformed between the first polymeric sheet and the second polymeric sheet;and a plurality of first perforations being formed through a portion ofthe first polymeric sheet that overlies the first sparging area so thatgas can pass from the first sparging area, through the firstperforations and into the chamber of the bag.
 2. The container assemblyas recited in claim 1, wherein the first weld line bounds a perimeter ofa first sparger pathway formed between the first polymeric sheet and thesecond polymeric sheet, the first sparger pathway comprising a first gastransfer path extending from a first end to an opposing second end andthe first sparging area formed at the second end and communicating withthe first gas transfer path.
 3. The container assembly as recited inclaim 2, wherein the first gas transfer path is constricted relative tothe first sparging area.
 4. The container assembly as recited in claim2, wherein the first gas transfer path is elongated while the firstsparging area is substantially circular.
 5. The container assembly asrecited in claim 2, wherein there are no perforations extending throughthe portion of the first polymeric sheet that overlies the first gastransfer path.
 6. The container assembly as recited in claim 2, furthercomprising a first gas line coupled with the first end of the first gastransfer path.
 7. The container assembly as recited in claim 1, furthercomprising a first gas line welded between the first polymeric sheet andthe second polymeric sheet.
 8. The container assembly as recited inclaim 1, further comprising a hole formed on the second polymeric sheet,a port secured to the second polymeric sheet and extending through thehole, and a first gas line coupled with the port.
 9. The containerassembly as recited in claim 1, further comprising a hole formed on thefirst polymeric sheet, a port secured to the first polymeric sheet andextending through the hole, and a first gas line coupled with the port.10. The container assembly as recited in claim 1, further comprising: ansecond weld line welding the first polymeric sheet to the secondpolymeric sheet, the second weld line bounding a second sparging areaformed between the first polymeric sheet and the second polymeric sheet;and a plurality of second perforations being formed through a portion ofthe first polymeric sheet that overlies the second sparging area. 11.The container assembly as recited in claim 10, further comprising amanifold in fluid communication with the first sparging area and thesecond sparging area.
 12. The container assembly as recited in claim 11,wherein the manifold comprises: a first valve that regulates the flow ofgas through the manifold and to the first sparging area; and a secondvalve that regulates the flow of gas through the manifold and to thesecond sparging area.
 13. The container assembly as recited in claim 12,wherein the first valve and the second valve are electrically operated.14. The container assembly as recited in claim 12, wherein the firstvalve and the second valve each comprise a hose clamp.
 15. The containerassembly as recited in claim 11, wherein the manifold comprises: a bodywith a gas inlet port and a plurality of gas outlet port, the gas inletport being in fluid communication with each of the plurality of gasoutlet ports; a first gas line being in fluid communication with thefirst sparging area and a first one of the gas outlet ports; and asecond gas line being in fluid communication with the second spargingarea and a second one of the gas outlet ports.
 16. The containerassembly as recited in claim 10, wherein the first perforations have adifferent size or number than the second perforations.
 17. The containerassembly as recited in claim 1, further comprising a rigid supporthousing bounding a compartment, the flexible bag being disposed withinthe compartment of the support housing.
 18. A container assemblycomprising: a flexible bag having an interior surface bounding a chamberand an opposing exterior surface, the bag having a bottom end wall beingcomprised of a first polymeric sheet overlying a second polymeric sheet;an first weld line welding the first polymeric sheet to the secondpolymeric sheet, the first weld line bounding a first sparging areaformed between the first polymeric sheet and the second polymeric sheet,a plurality of first perforations being formed through a portion of thefirst polymeric sheet that overlies the first sparging area; and ansecond weld line welding the first polymeric sheet to the secondpolymeric sheet, the second weld line bounding a second sparging areaformed between the first polymeric sheet and the second polymeric sheet,a plurality of second perforations being formed through a portion of thefirst polymeric sheet that overlies the second sparging area.
 19. Thecontainer assembly as recited in claim 18, further comprising a manifoldin fluid communication with the first sparging area and the secondsparging area.
 20. The container assembly as recited in claim 19,wherein the manifold comprises: a first valve that regulates the flow ofgas through the manifold and to the first sparging area; and a secondvalve that regulates the flow of gas through the manifold and to thesecond sparging area.
 21. The container assembly as recited in claim 20,wherein the first valve and the second valve are electrically operated.22. The container assembly as recited in claim 20, wherein the firstvalve and the second valve each comprise a hose clamp.
 23. The containerassembly as recited in claim 19, further comprising: a first portcoupled with the second polymeric sheet and communicating with the firstsparging area; and a second port coupled with the second polymeric sheetand communicating with the second sparging area.
 24. The containerassembly as recited in claim 23, wherein the manifold comprise: a bodywith a gas inlet port and a plurality of gas outlet ports, the gas inletport being in fluid communication with each of the plurality of gasoutlet ports; a first gas line extending from the first port to a firstone of the gas outlet ports; and a second gas line extending from thesecond port to a second one of the gas outlet ports.
 25. The containerassembly as recited in claim 18, wherein the first perforations have adifferent size or number than the second perforations.
 26. A containerassembly comprising: a flexible bag having an interior surface boundinga chamber and an opposing exterior surface, the bag having a bottom endwall comprised of a first polymeric sheet overlying a second polymericsheet; at least one weld line welding the first polymeric sheet to thesecond polymeric sheet, the at least one weld line bounding an elongatedsparging area formed between the first polymeric sheet and the secondpolymeric sheet and extending between a first end and an opposing secondend, a plurality of perforations being formed through a portion of thefirst polymeric sheet that overlies the first sparging area; a first gasline coupled with the first end of the sparging area; and a second gasline coupled with second end of the sparging area.
 27. A containerassembly as recited in claim 26, a manifold coupled with the first gasline and the second gas line.
 28. A container assembly as recited inclaim 26, wherein the sparging area covers at least 40% of the surfacearea of one side of the bottom end wall.
 29. The container assembly asrecited in claim 26, further comprising a rigid support housing boundinga compartment, the flexible bag being disposed within the compartment ofthe support housing.